Issue 25, 2016

Label-free DNA hairpin probe for real-time monitoring of alkaline phosphatase activity

Abstract

We have developed a new methodology for the label-free detection of alkaline phosphatase (ALP) activity based on a DNA hairpin and thioflavin T (ThT) probes. In the presence of ALP, the 3′-phosphate of the primer DNA is dephosphorylated, triggering a primer extension reaction to open the hairpin probe and leading to release of the G-quartets. These then bind to ThT to form ThT/G-quadruplexes with obvious fluorescence generation. The method is highly sensitive, and the ALP detection limit was estimated to be 0.001 U mL−1, which is superior or comparable to those of reported assays. The assay avoids complicated modifications or chemical labeling, making it simple and cost-effective. Furthermore, the inhibition of ALP activity by the inhibitor theophylline is demonstrated, indicating the potential for the discovery and characterization of ALP-targeted drug candidates in pharmaceutical development.

Graphical abstract: Label-free DNA hairpin probe for real-time monitoring of alkaline phosphatase activity

Article information

Article type
Paper
Submitted
05 Apr 2016
Accepted
27 May 2016
First published
01 Jun 2016

Anal. Methods, 2016,8, 5095-5100

Label-free DNA hairpin probe for real-time monitoring of alkaline phosphatase activity

J. Du, L. Xiong, C. Ma, H. Liu, J. Wang and K. Wang, Anal. Methods, 2016, 8, 5095 DOI: 10.1039/C6AY00989A

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